Fibroblast Migration Increased by PEG-EGF in Hydrogels

1
Fibroblast Migration Increased by PEG-EGF in
Hydrogels

• Student Name
• Date

2
Objective

• Does epidermal growth factor (EGF) tethered to
  PEG increase cell number and migration of
  fibroblasts in hydrogels?

3
Measuring Cell Proliferation

• Hydrogels with GGLG-PAGGK-PEG were prepared with
  or without PEG-EGF
• 20,000 cells placed on media-soaked gel and
  incubated 48 hours
• PCNA stained cells were counted
• Control performed similarly with liquid media and
  cell number determined with Coulter Counter

4
Measuring Cell Migration

• Hydrogels on membranes were prepared containing
  either PEG-EGF or PEG-RGDS, both PEG-EGF and
  PEG-RGDS, or neither
• 50,000 cells were added and incubated for 7 days
• Calculated migration index as of cells that
  move through membrane as determined by Coulter
  Counter

5
EGF Increases Cell Number

• More cells in presence of EGF
• Mean/-s.d. n6
• 20,000 initial cell number
• p0.07 vs. no EGF

- toxin
toxin
Number of cells
PEG-EGF
EGF
No EGF
6
EGF Increases Number of Cells in S-Phase
- toxin

• More cells in S-phase with EGF
• Mean/-s.d. n6
• p0.05 vs. no EGF

toxin
Percentage of cells
PEG-EGF
EGF
No EGF
7
EGF in Hydrogels

• EGF increases number of cells above 20,000
  initial cell number (shown as dashed line)
• Mean/-s.d. n7
• p0.05 vs. no EGF

35,000


30,000
25,000
20,000
Number of cells
15,000
10,000
5,000
0
PEG-EGF
EGF
No EGF
8
Migration occurs with RGDS and EGFRGDS

• Migration occurs with RGDS and RGDSEGF
• Migration doesnt occur with EGF alone
• Migration occurs in presence of toxin

- toxin
toxin
Mean /- s.d. n4
Number of cells
EGF RGDS
PEG
EGF
RGDS
9
Fibroblast Migration Increased by PEG-EGF

• PEG-EGF increases cell number and percent of
  S-phase cells
• PEG-EGF with PEG-RGDS causes migration through
  hydrogel

10
Data and results taken from the paper Effects of
Epidermal Growth Factor on Fibroblast Migration
through Biomimetic Hydrogels, A.S. Gobin, J.L.
West, Biotechnol. Prog. 2003, 19, 1781-1785. Dr.
A. Saterbak accepts all responsibility for any
misrepresentation of any part of the paper.
Poster developed entirely for pedagogical purpose
for BIOE342.